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Mouse clinic - transgenic core facility

The Netherlands Cancer Institute has a very strong history in the generation and validation of mouse models of human cancer. Many conditional mouse strains for tumor suppressor alleles often lost or mutated in human cancer, such as Trp53, PTEN and Cdkn2a, have been generated here and are routinely used worldwide. In that context, the transgenic core facility accommodates all activities required to generate genetically modified animals, from DNA and Embryonic Stem Cell (ESC) services to transgenic services, such as ESC microinjection, zygote injection as well as cryopreservation. State-of-the-art technologies, such as ESC re-derivation from complex models and genome editing, are also offered.

The transgenic core facility is part of the Mouse Clinic for Cancer and Aging (MCCA). The MCCA is a collaboration of the Netherlands Cancer Institute and the European Research Institute for the Biology of Ageing in Groningen, The Netherlands. The MCCA is funded by a Roadmap grant from the Netherlands Organisation for Scientific Research (NWO).

Websites: www.mccanet.nl or gem.nki.nl

Services

Generating genetically engineered mice is a complex process that requires a good strategy and detailed planning. In consultation with the researcher we provide advice on the strategy and construct design for generating the mouse strain tailor-made to answer a biological question. Depending on the complexity and workload a practical plan is devised to reach that goal and in some case might involve participation of the researcher in cloning and/or ESC culture. Our services are listed below and include cryopreservation of new and existing strains.

DNA services:

  • Generation of targeting and Flp-in constructs
  • Design screening PCRs for targeting constructs
  • Design and cloning of CRISPR gRNA plasmids
  • Design ssODN for homology directed repair

ESC services:

  • ESC re-derivation and cryogenic storage
  • ESC quality testing
  • Targeting and Flp-in of ESCs
  • PCR and Southern blot screening

Transgenic services:

  • ESC micro-injection/generation of chimeric mice
  • Zygote injection; RNA and/or DNA injection
  • Cryopreservation - sperm
  • Cryopreservation - embryo

Publications

Semenova EA, Kwon M-C, Monkhorst K, Song JY, Bhaskaran R, Krijgsman O, Kuilman T, Peters D, Buikhuisen WA, Smit EF, Pritchard C, Cozijnsen M, van der Vliet J, Zevenhoven J, Lambooij JP, Proost N, van Montfort E, Velds A, Huijbers IJ, Berns A. Transcription factor NFIB is a driver of Small Cell Lung Cancer progression in mice and marks metastatic disease in patients. Cell Reports. 2016;16:631-43.

Annunziato S, Kas SM, Nethe M, Yücel H, Del Bravo J, Pritchard C, Bin Ali R, van Gerwen B, Siteur B, Drenth AP, Schut E, van de Ven M, Boelens MC, Klarenbeek S, Huijbers IJ, van Miltenburg MH, Jonkers J. (2016) Modeling invasive lobular breast carcinoma by CRISPR/Cas9-mediated somatic genome editing of the mammary gland. Genes Dev. 30:1470-80.

Pilzecker B, Buoninfante OA, Pritchard C, Blomberg OS, Huijbers IJ, van den Berk PC, Jacobs H. (2016) PrimPol prevents APOBEC/AID family mediated DNA mutagenesis. Nucleic Acids Res. 44: 4734-44.

Huijbers IJ, Del Bravo J, Bin Ali R, Pritchard C, Braumuller TM, van Miltenburg MH, Henneman L, Michalak EM, Berns A & Jonkers J. (2015) Using the GEMM-ESC strategy to study gene function in mouse models. Nature Protocols 10: 1755-85.

Henneman L, van Miltenburg MH, Michalak EM, Braumuller TM, Jaspers JE, Drenth AP, de Korte-Grimmerink R, Gogola E, Szuhai K, Schlicker A, Bin Ali R, Pritchard C, Huijbers IJ, Berns A, Rottenberg S, Jonkers J. (2015) Selective resistance to the PARP inhibitor olaparib in a mouse model for BRCA1-deficient metaplastic breast cancer. PNAS 2015 Jun 22.

Bin Ali R, van der Ahé F, Braumuller TM, Pritchard C, Krimpenfort P, Berns A, Huijbers IJ. (2014) Improved pregnancy and birth rates with routine application of nonsurgical embryo transfer. Transgenic Res. 23: 691-5.

Huijbers IJ, Bin Ali R, Pritchard C, Cozijnsen M, Kwon MC, Proost N, Song JY, de Vries H, Badhai J, Sutherland K, Krimpenfort P, Michalak EM, Jonkers J, Berns A (2014) Rapid target gene validation in complex cancer mouse models using re-derived embryonic stem cells. EMBO Mol Med 6: 212-25.
 
Huijbers IJ, Krimpenfort P, Berns A and Jonkers J. (2011) Rapid validation of cancer genes in chimeras derived from established genetically engineered mouse models. Bioessays 33, 701-710

People working at the Mouse clinic - transgenic core facility

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Ivo Huijbers

Head transgenesis unit

Personal details

Experience

Since October 2012, I coordinate the embryonic stem cell (ESC) and transgenic activities of the Mouse Clinic for Cancer and Aging research. I provide consultancy in the design of desired transgenic models and lead a team of technicians that performs ESC re-derivation, culture and manipulation and that generates new mouse strains by ESC injection.
My interest in generating mouse models originates from my PhD at the LICR in Brussels, where I made knockout and transgenic mice to study immunotherapy. After 4 years of postdoctoral research in London (ICR/Breakthrough Breast Cancer Research Centre), I returned to the Netherlands (group of Prof. Anton Berns) to setup a method for the fast generation of cancer mouse models with re-derived ESCs.

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Rahmen Bin Ali

Technician

Personal details

Experience

Originally from Singapore, I have been working at the department of Molecular Genetics and in particular in the transgenic facility since 2002. My job involves the practical workflow in a transgenic facility, focusing mainly on the making of transgenic and/or knockout mice as a cancer model. In addition, I am responsible for the running of the embryonic stem cell (ES) lab and in that context re-derive ES cells from complex mouse models present within the institute. Recently, I am also involved with the mouse sperm cryopreservation, mouse embryos vitrification and IVF.

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Tanya Vermeeren-Braumuller, BASc

Technician

Experience

In 2006 I did my internship in the lab of Jos Jonkers at the Netherlands Cancer Institute. In 2007 I obtained my Bachelor of Applied Sciences (Hogeschool Leiden) and started working as a technician at the Netherlands Cancer Institute. Since 2012 I have a joint appointment in the group of Jos Jonkers and the transgenics facility headed by Ivo Huijbers. I perform embryonic stem (ES) cell derivations, ES cell culture and blastocyst injections. I like working at the NKI, because I get a lot of responsibility and there is a good collaboration between the colleagues of our lab.

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Linda Henneman, Ph.D.

Technician

Experience

I finished my Master in Biomedical Science at the VU University Amsterdam in 2004. In the same year I started as a PhD student in the group of Ron Wanders at the Academic Medical Centre in Amsterdam. I received my PhD in 2011. In 2009, I started as a postdoc in the groups of Ed Roos and Kees Jalink at the NKI. In 2011, I started my second postdoc in the group of Jos Jonkers, where I developed genetically engineered mouse models (GEMMs) to study ER-positive breast cancer. I also used GEMMs to validate several candidate oncogenes in BRCA1-associated breast cancer. In 2016, I joined the transgenic facility.

 

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Colin Pritchard

Technician

Experience

Recently some significant technological advances have taken place that facilitate and extend the possibilities to manipulate the mouse genome. In our transgenic facility we aim to apply, mould and direct this knowledge in ways that maximizes its useful implementation in cancer research. In this context I work mostly on technology development rather than the already applied offerings of the transgenic facility. 

My work currently includes:

  • The development, design, cloning and testing of DNA constructs
  • The application and adaptation of new tools (e.g. CRISPRs)
  • The derivation and manipulation of ES cells
  • Blastocyst injection
  • Discuss experimental possibilities with users of the facility.
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Fina van der Ahé

Technical Staff

Experience

Since 2001 I have been working as a biotechnician in the group of Prof. Anton Berns and in the Mouse clinic transgenesis unit. My job involves microinjection, genotyping, and sperm cryopreservation.

 

 

 

 

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Kroese, Lona

Technician

Experience

Fresh from university, this is my first real job. I gained my lab experience through several internships either as a bachelor student in Medical Natural Science or as a master student in Cardiovascular Research. As a technician at the MCCA I will contribute to the development of genetically engineered mouse models by introducing genetic modifications in embryonic stem cells.  I expect to learn a lot and want to develop myself to be of great support to this group.

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Paul Krimpenfort

Academic Staff

Experience

My main scientific interest is the tumor suppressive function of the Ink4 gene family encoding the Cdk4/6 inhibitors p15Ink4b, p16Ink4a, p18Ink4c and p19Ink4d. Since Cdk4 and Cdkn6 stimulate G1 progression by phosphorylating Rb, the primary effect of Ink4 inactivation is thought to be the loss of cell cycle control. However, apart from their role in cell cycle control recent observations suggest the implication of the Ink4 proteins in tumor cell behaviour e.g. migration and invasiveness. An important issue is if and to what extent the Ink4 genes are redundant.

In addition, I am also involved in the NKI Transgenic Facility and I am a member of the Animal Experiment Committee (DEC) of the NKI.

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